Abstract: The overall goal of this study is to develop and demonstrate a novel, predictive in vitro organotypic 3D-culture models of the blood-brain barrier (BBB) derived from rodents for chemical toxicity screening. In contrast with current Transwell based static and microfluidic assays, our commercially available SynBBB model enables real-time visualization and quantitation of transport/permeation under physiological microcirculatory size and flow conditions, while simultaneously simplifying on chip and off chip analysis. The apical side provides a 3D architecture of endothelial cells while the basolateral side supports 3D glial cell co-cultures. Phase I study seeks to demonstrate the feasibility of the SynBBB model for predicting central nervous system toxicity of essential and heavy metals specifically across the BBB. It will culminate with a clear demonstration of the model with validation against in vivo studies. During Phase II, we will develop a higher throughput platform (12-24 assays) with automated measurement of physiological end-points for mechanistic understanding and detailed validation against animal studies. A multi-disciplinary, industry-academic partnership including CFDRC and Albert Einstein College of Medicine encompassing expertise in microfluidics cell-based assays, blood brain barrier, chemical and heavy metal derived neurotoxicity, drug discovery and development, and therapeutic evaluation will oversee the development of this model. The developed assay will have critical applications in basic research for understanding of chemical toxicity mechanisms and development of toxin neutralization strategies. The end-product will be commercialized to government agencies, pharmaceutical firms, drug research labs and universities/non-profit centers engaged in chemical toxicity screening, drug discovery, and drug delivery.